ANAC-C (PR3) hs ELISA Assay Kit

$575.00

The Eagle Biosciences ANAC-C (PR3) hs ELISA Assay Kit is intended for the quantification of IgG auto-antibodies against Proteinase 3 in serum or plasma.The ANAC-C (PR3) hs ELISA Assay Kit is for research use only and should not be used for diagnostic procedures.

SKU: ARG80360 Categories: , ,

ANAC-C (PR3) hs ELISA Assay Kit

For Research Use Only

Size: 1 x 96 wells
Sensitivity: 0.5 µg/ml
Dynamic Range: 10-200 µg/ml
Incubation Time: 1 hour
Sample Type: Serum, Plasma
Sample Size: 5 µl

Additional Information

Assay Principle

The Eagle Biosciences ANAC-C (PR3) hs ELISA Assay Kit is an Enzyme Immunoassay for the quantification of IgG auto-antibodies against Proteinase 3 in serum or plasma.  This assay employs the quantitative enzyme immunoassay technique. A highly purified Proteinase 3 (PR3) has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any PR3 antibodies (Ab) present is bound by the immobilized antigen. After washing away any unbound substances, a HRP-conjugated anti-human antibody is added to each well and incubate. After washing away any unbound antibody-enzyme reagent, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of Ab bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450nm ±2nm.The concentration of PR3 Ab in the sample is then determined by comparing the O.D of samples to the standard curve.

Assay Procedure

  1. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it.
  2. Add 100µl of standards, controls, samples and zero controls into wells.
  3. Incubate for 30 minutes at RT.
  4. Aspirate each well and wash, repeating the process 4 times for a total 5 washes. Wash by filling each well with 1× Wash Buffer (350µl) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating, decanting or blotting against clean paper towels.
  5. Add 100µl 1X HRP-Antibody Conjugate into each well. Incubate for 15 minutes at RT.
  6. Wash as according to step 4.
  7. Add 100µl of TMB Reagent to each well. Incubate for 15 minutes at room temperature.
  8. Add 100µl of Stop Solution to each well. Incubate for 5 minutes at RT. The color of the solution should change from blue to yellow.
  9. Read the OD with a microplate reader at 450nm immediately.

Quality Assurance

Intra-assay and Inter-assay precision. The CV value of intra-assay precision was 2.6% and inter-assay precision was 4%.

Manual

Product Manual